TWI770565B - Methods for selecting a drug, and uses of the drug for treating psoriasis - Google Patents

Abstract

Provided herein are directed to methods for selecting a drug for treating psoriasis in a subject. Accordingly, also encompassed within the present disclosure are methods for treating psoriasis, with the aid of said drug in treating a subject afflicted with psoriasis.

Description

A method for screening a drug, and the use of the drug in the treatment of psoriasis

The present disclosure generally relates to the technical field of psoriasis treatment. Specifically, the present disclosure relates to a drug screening method for screening a drug for treating psoriasis from a plurality of candidate drugs, and a method for treating psoriasis by using the therapeutic drug.

Psoriasis is a chronic inflammatory skin disease mediated by immunity. It is generally believed that psoriasis is related to immune dysfunction and genetics. However, other factors such as seasonal changes, stress, viruses, bacteria, metabolic disorders, drug-induced, Trauma and diet are also considered to have the potential to induce psoriasis. The typical symptoms of psoriasis are lumpy, abnormal skin lesions that appear on the surface of the patient's skin, with redness, itching, and scaling in the affected area. The prevalence of psoriasis is known to be approximately 2-4%, women and men are equally affected, and the prevalence is higher in adults (approximately 0.91%) than children (approximately 0-2.1%). -8.5%).

Current treatments for psoriasis include topical therapy, phototherapy, photochemotherapy or systemic therapy. For topical therapy, drugs such as steroid, Calcipotriol, Anthralin, or Coal tar are used locally on the affected area. Phototherapy uses ultraviolet rays to irradiate the skin of patients. It is currently the first-line treatment for psoriasis. However, patients often need to go to a treatment center with phototherapy equipment for treatment, which is troublesome, and ultraviolet rays also have potential carcinogenic risks. In the part of photochemotherapy, it refers to the combined therapy of oral psoralen and ultraviolet irradiation with a wavelength of 320-400 nm. As for systemic therapy, biological agents (eg, therapeutic antibodies) that can treat psoriasis are used, or cyclosporine (Cyclosporine), methotrexate (Methotrexate), or tretinoin (Tretinoin).

Although the above-mentioned various methods have been used to treat psoriasis, it is often found in clinical practice that because the individual physiques of individual patients are different, it is not easy for clinicians to find a suitable treatment for individual patients from the above treatments. As for the poor response of patients to inappropriate therapy, the patient's compliance with treatment is reduced, and medical resources are wasted.

In view of this, in order to improve the accuracy of medication for psoriasis patients and achieve the realm of personalized medicine, it is urgent in related fields to develop a method that can improve the precise medication of psoriasis, so as to improve the effectiveness and safety of medication for patients. , and reduce the waste of medical resources.

A brief summary of the present disclosure is presented below to facilitate the reader to have a basic understanding of the present disclosure. This summary is not an extensive overview of the disclosure, nor is it intended to identify key/critical elements of the disclosure or to delineate the scope of the disclosure. Its sole purpose is to present some concepts of the disclosure in a simplified form as a prelude to the more detailed description that is presented later.

Accordingly, the purpose of the present disclosure is to provide a drug screening method, so as to select a therapeutic drug suitable for an individual psoriasis patient from a plurality of candidate drugs, so as to treat the psoriatic disease of the patient. As embodied and broadly described in this disclosure, one aspect of this disclosure relates to a method of screening a therapeutic drug from a plurality of drug candidates, wherein the therapeutic drug is for treating a patient suffering from psoriasis the individual. The method includes: (a) Obtain an isolated peripheral blood mononuclear cell (PBMC) from the individual; (b) Activating the peripheral blood mononuclear cells of step (a) with a stimulus to make the cells produce interferon-gamma (IFN-γ) and interleukin-13 (interleukin-13, IL-13) ; (c) respectively administering the plurality of candidate drugs to the activated peripheral blood mononuclear cells in step (b); (d) respectively detecting the expression levels of the interferon-γ and the interleukin-13 in the peripheral blood mononuclear cells of step (c); and (e) Based on the expression levels of the interferon-γ and the interleukin-13 detected in step (d), the therapeutic drug is screened from the plurality of candidate drugs, wherein the therapeutic drug is the interferon-γ The expression amount of γ has the highest inhibitory ability, or the ratio of the expression amount of interferon-γ to the expression amount of interleukin-13 is the smallest.

According to an embodiment of the present disclosure, the therapeutic drug reduces the amount of interferon-gamma expression by at least 1% after administration of the candidate drug compared to before administration of the candidate drug. According to another embodiment of the present disclosure, wherein the therapeutic drug is at least 1% lower in the expression of interferon-gamma and interleukin-13 after administration of the candidate drug than before administration of the candidate drug Performance ratio.

According to certain embodiments of the present disclosure, the irritant is a Gram-positive bacteria, a Gram-negative bacteria, or a lipopolysaccharide (LPS). According to an operational embodiment of the present disclosure, the gram-positive bacteria Streptococcus pyogenes . According to other embodiments of the present disclosure, the Gram-negative bacteria is Escherichia coli .

According to certain embodiments of the present disclosure, each of the plurality of drug candidates is selected from the group consisting of: Acitretin, alefacept, Anthralin, Apremilast (Acitretin). Apremilast), Betamethasone, Calcipotriene, Calcipotriol, Calcitriol, Clobetasol, Coal tar, Cyclosporine ( Cyclosporine, Dithranol, Etanercept, Fluocinolone, Hydrocortisone, Infliximab, Methotrexate, Pyridoxine Pimecrolimus, Tacrolimus, Tazarotene, Tretinoin, ALX-0761, BCD-085, Bimekizumab, Bodalumab (Brodalumab), CJM112, CNTO 6785, COVA322, Ixekizumab, LY3114062, MSB0010841, NI-1401, Perakizumab, Remtolumab, RG7624, Secuqi Secukinumab, Vunakizumab, Brazikumab, Guselkumab, Mirikizumab, Risankizumab , Tildrakizumab, Ustekinumab, Adalimumab, Certolizumab, Etanercept, Golimumab ( Golimumab), Infliximab (Infliximab), and lenercept (Lenercept). According to a specific embodiment, the plurality of drug candidates are obtained from: adalimumab, golimumab, guselkumab, ixelizumab, secukinumab, and ustekinumab. composition.

Another aspect of the present disclosure pertains to a method for treating an individual suffering from psoriasis. The method includes: (1) Screen a therapeutic drug from a plurality of candidate drugs by the following steps, wherein the therapeutic drug is used to treat the psoriasis of the individual, including: (1a) Obtaining an isolated peripheral blood mononuclear cell from the individual; (1b) activate the peripheral blood mononuclear cells of step (1a) with a stimulant, so that the cells produce interferon-γ and interleukin-13; (1c) respectively administering the plurality of candidate drugs to the activated peripheral blood mononuclear cells in step (1b); (1d) respectively detecting the expression levels of the interferon-γ and the interleukin-13 in the peripheral blood mononuclear cells of step (1c); and (1e) Based on the expression levels of the interferon-γ and the interleukin-13 detected in step (1d), the therapeutic drug is selected from the plurality of candidate drugs, wherein the therapeutic drug is the interferon-γ The expression of gamma has the highest inhibitory power, or the ratio of the expression of interferon-gamma to the expression of interleukin-13 is the smallest; and (2) Administer an effective amount of the therapeutic drug of step (1e) to the individual.

According to an embodiment of the present disclosure, the therapeutic drug reduces the amount of interferon-gamma expression by at least 1% after administration of the candidate drug compared to before administration of the candidate drug. According to another embodiment of the present disclosure, wherein the therapeutic drug is at least 1% lower in the expression of interferon-gamma and interleukin-13 after administration of the candidate drug than before administration of the candidate drug Performance ratio.

According to certain embodiments of the present disclosure, the irritant is a gram-positive bacteria, a gram-negative bacteria, or a lipopolysaccharide. In a preferred embodiment, the Gram-positive bacteria are Streptococcus pyogenes. In another preferred embodiment, the Gram-negative bacteria is Escherichia coli.

According to certain embodiments of the present disclosure, each of the plurality of drug candidates is selected from the group consisting of: Acitretin, Afacept, Allionol, Apremilast, Betamethasone, Calcipotriene , calcipotriol, calcitriol, becloxone, tar, cyclosporine, diconitol, etanercept, fluocinolone, hydrocortisone, infliximab, ametrexate, pimecropin Limus, tacrolimus, tazarotene, retinoic acid, ALX-0761, BCD-085, pembrolizumab, bodalumab, CJM112, CNTO 6785, COVA322, ixelizumab, LY3114062, MSB0010841, NI-1401, perelizumab, rendolizumab, RG7624, secukinumab, vonazumab, blaculumab, guselkumab, milizumab, rukinizumab Salizumab, tiltaximab, ustekinumab, adalimumab, certolizumab, etanercept, golimumab, infliximab, and lenacept formed groups. In a specific embodiment, the plurality of drug candidates are composed of: adalimumab, golimumab, guselkumab, ixelizumab, secukinumab, and ustekinumab composed.

According to certain embodiments of the present disclosure, an individual suitable for use in the methods of the present disclosure to treat psoriasis is a human.

Numerous attendant features and advantages of the present disclosure will become readily apparent upon review of the following detailed description and accompanying drawings.

In order to make the description of the present disclosure more detailed and complete, the following provides an illustrative description for the embodiments and specific embodiments of the present invention; but this is not the only form of implementing or using the specific embodiments of the present invention. The features of various specific embodiments as well as method steps and sequences for constructing and operating these specific embodiments are encompassed in the detailed description. However, other embodiments may also be utilized to achieve the same or equivalent function and sequence of steps.

i. definition

For convenience, specific terminology used in the description, the embodiments and the appended claims are collected here. Unless otherwise defined in this specification, scientific and technical terms used herein have the same meanings as understood and commonly used by those of ordinary skill in the art to which this invention belongs. Moreover, without conflicting with the context, the singular noun used in this specification covers the plural form of the noun; and the plural noun used also covers the singular form of the noun. Specifically, in this specification and the scope of the claims, the singular form "a" (a and an) includes plural references unless the context dictates otherwise. In addition, in this specification and the scope of the patent application, expressions such as "at least one" and "one or more" have the same meaning, and both mean that one, two, three or more are included. What's more, in this specification and the scope of the patent application, "at least one of A, B and C", "at least one of A, B or C" and "at least one of A, B and/or C" ” means A only, B only, C only, both A and B, B and C, and C, and A, B, and C.

Notwithstanding that the numerical ranges and parameters setting forth the broader scope of the invention are approximations, the numerical values set forth in the specific examples are reported as precisely as possible. Any numerical value, however, inherently contains the standard deviation resulting from individual testing methods. As used herein, the term "about" generally refers to an actual value within plus or minus 10%, 5%, 1%, or 0.5% of a particular value or range. Alternatively, the word "about" means that the actual value lies within an acceptable standard error of the mean, as considered by one of ordinary skill in the art to which this invention pertains. Except for experimental examples, or unless otherwise expressly stated, all ranges, quantities, values and percentages used herein (eg, to describe material amounts, time durations, temperatures, operating conditions, quantity ratios and others) should be understood similar) are modified by "about". Therefore, unless otherwise stated to the contrary, the numerical parameters disclosed in this specification and the accompanying claims are approximate numerical values and may be changed as required. At a minimum, these numerical parameters should be construed to mean the number of significant digits indicated and the numerical values obtained by applying ordinary rounding. Numerical ranges are expressed herein as being from one endpoint to the other or between the endpoints; unless otherwise indicated, the numerical ranges recited herein are inclusive of the endpoints.

The term "treatment" or "treating" as used herein may refer to a curative or palliative measure. Specifically, as used herein, the term "treatment" refers to the administration or administration of an effective amount of a therapeutic agent of the present disclosure to a subject, the therapeutic agent being screened using the drug screening methods of the present disclosure, and The individual suffers from psoriasis, suffers from symptoms associated with psoriasis, a disease or disorder of psoriasis whereby partial or complete alleviate, ameliorate, relieve, Delay onset, inhibit progression, reduce severity, and/or reduce the incidence of one or more symptoms or features of psoriasis.

As used herein, the term "activate" (activate, activating, or activation) refers to the use of a specific substance (eg, streptococcus) to stimulate a living body or a living cell (eg, peripheral blood mononuclear cells), so that the living body or Live cells induce a corresponding immune response (eg, secrete related inflammatory cytokines, such as: interferon-gamma, interleukin-13, interleukin-6, interleukin-8) , interleukin-12, or interleukin-17, etc.).

As used herein, the terms "administered," "administering," or "administration" are used interchangeably herein and refer to the direct contact of a target (eg, peripheral blood mononuclear cells) with a specific substance (eg, the present disclosure). The act of administering a specific substance (eg, a therapeutic drug of the present disclosure) to an individual (eg, a human). For example, a drug candidate of the present disclosure is directly added to a culture dish containing peripheral blood mononuclear cells for co-cultivation for a period of time. Furthermore, when a therapeutic agent of the present disclosure is administered to an individual, it is meant via oral, intracranial, intraspinal, intrathecal, intramedullary, intracerebral, intracerebroventricular, intravenous, intraarterial, intracardiac, intradermal, subcutaneous , transdermal, intraperitoneal, or intramuscular routes to administer the therapeutic agents of the present disclosure to the individual in need thereof.

As used herein, the term "an effective amount" refers to an amount effective to achieve the desired therapeutic effect (eg, to treat psoriasis) in the dose and time period necessary for treatment of the therapeutic agents of the present disclosure. dosage of use. For therapeutic purposes, an effective amount also means that the therapeutic benefits of a component of a drug outweigh the toxic or detrimental effects of that component. An effective amount of an agent does not necessarily cure the disease or disorder, but delays, retards or prevents the development of the disease or disorder, or alleviates the symptoms associated with the disease or disorder. An effective amount can be divided into one, two or more doses and administered one, two or more times over a specified period in the appropriate dosage form. The specific effective amount depends on a variety of factors, for example, the condition to be treated, the physiological condition of the individual (eg, the individual's weight, age, or sex), the species being treated, the duration of the treatment, concomitant therapies (if any) ), and the specific dosage form used, and the structure of the compound or its derivatives. An effective amount may be expressed in any suitable manner. For example, an effective amount of an agent can be expressed as the total weight of the drug (eg, grams, milligrams, or micrograms), or as a ratio of the weight of the drug to body weight (eg, milligrams per kilogram of body weight (mg/kg, mg/kg) Kg)). Alternatively, an effective amount of an agent can be expressed as a concentration, eg, molar concentration, mass concentration, volume concentration, molality, molar fraction ( mole fraction, mass fraction and mixing ratio. Appropriate doses range from 0.01 mg to 100.0 mg per kilogram of body weight. As will be appreciated, a wide range of adjustments in the desired dosage are also contemplated, depending upon the different compositions and the different efficacies resulting from the different routes of administration. For example, oral administration is expected to require higher doses than intravenous injection. Those skilled in the art fully understand that the dosage can be adjusted according to rules of thumb. One of skill in the art can convert doses obtained based on experimental animal models into human equivalent doses (HEDs) of drugs (eg, therapeutic drugs of the present disclosure). For example, those skilled in the art can use the "Estimating the Maximum Safe Starting Dose of Adult Healthy Volunteers in Initial Clinical Treatment Tests" published by the US Food and Drug Administration (FDA) (U.S. Food and Drug Administration, FDA). Safe Starting Dose in Initial Clinical Trials for Therapeutics in Adult Healthy Volunteers) to estimate the highest safe dose for human use.

As used herein, the terms "subject" or "patient" are used interchangeably to refer to an animal, including humans, that can be treated with a therapeutic agent of the present disclosure, wherein the therapeutic agent refers to a Those screened by the screening method of the present disclosure. Unless one of the genders is specified, the terms "individual" and "patient" refer to both males and females. Accordingly, the terms "individual" and "patient" include any mammal that would benefit from treatment with the therapeutic agents of the present disclosure. Examples of "individuals" and "patients" that can be treated with therapeutic agents of the present disclosure include, but are not limited to, humans, rats, mice, guinea pigs, monkeys, pigs, goats, cows, horses, dogs, cats, Birds and Chickens. In an exemplary embodiment, the individual is a mouse. In another illustrative embodiment, the individual is a human.

The term "pharmaceutically acceptable" refers to molecular entities and compositions that are "generally regarded as safe", eg, that are physiologically tolerable and In humans, allergic reactions or similar adverse reactions, such as nausea, dizziness, etc., generally do not occur. Preferably, the term "pharmaceutically acceptable" as used herein means approved by a regulatory agency of the federal or state government or listed in the US Pharmacopeia or other generally recognized pharmacopeia for use in animals, particularly is for humans.

II. Detailed description of the invention

The purpose of the present disclosure is to provide a method for screening a drug suitable for the treatment of psoriasis from a plurality of candidate drugs, and the method can screen out a psoriasis drug suitable for the patient according to the different constitutions of individual patients with psoriasis, thereby Improve the effectiveness and safety of psoriasis treatment, and reduce the waste of medical resources.

1. Methods of Screening Drugs

Accordingly, one aspect of the present disclosure relates to a method for screening a drug, wherein a therapeutic drug is selected from a plurality of candidate drugs, wherein the therapeutic drug is used to treat psoriasis in an individual, comprising: (a) Obtain an isolated peripheral blood mononuclear cell (PBMC) from the individual; (b) activating the peripheral blood mononuclear cells of step (a) with a stimulus, so that the cells produce interferon-γ (IFN-γ) and interleukin-13 (IL-13); (c) respectively administering the plurality of candidate drugs to the activated peripheral blood mononuclear cells in step (b); (d) respectively detecting the expression levels of the interferon-γ and the interleukin-13 in the peripheral blood mononuclear cells of step (c); and (e) Based on the expression levels of the interferon-γ and the interleukin-13 detected in step (d), the therapeutic drug is screened from the plurality of candidate drugs, wherein the therapeutic drug is the interferon-γ The expression amount of γ has the highest inhibitory ability, or the ratio of the expression amount of interferon-γ to the expression amount of interleukin-13 has the highest inhibitory ability.

First, in step (a), a whole blood sample is collected from a patient suffering from psoriasis, and peripheral blood mononuclear cells are isolated from the whole blood sample. Next, the obtained peripheral blood mononuclear cells are treated with a stimulus for a period of time (eg, 16 to 72 hours, such as: 16, 18, 24, 36, 48, 60 or 72 hours; preferably, 24 hours hours), so that the peripheral blood mononuclear cells are activated by the stimulation of the streptococcus and secrete interferon-gamma, interleukin-13, and other inflammatory cytokines (for example, interleukin-6, interleukin-6, interleukin-8, interleukin-12, or interleukin-17, etc.) (step (b)).

According to embodiments of the present disclosure, the irritant is a gram-positive bacteria, a gram-negative bacteria, or a lipopolysaccharide. Exemplary Gram-positive bacteria include, but are not limited to, Bacillus (eg, Bacillus cereus , Bacillus thuringiensis , Bacillus anthracis ); Listeria ( Listeria ) (eg, Listeria monocytogenes ); Staphylococcus (eg, Staphylococcus aureus ), Staphylococcus albus ( Staphylococcus albus ), Staphylococcus citrus ( Staphylococcus citreus ); Streptococcus (described below), Enterococcus (described below); and Clostridium (eg, Clostridium botulinum , Clostridium butyricum) ( Clostridium butyricum ), Clostridium difficile , Clostridium perfringens , Clostridium tetani ). In a preferred embodiment, the Gram-positive bacteria are Streptococcus.

According to one embodiment of the present disclosure, the Streptococcus is Group A Streptococcus , eg, Streptococcus pyogenes. According to another embodiment of the present disclosure, the Streptococcus is Group B Streptococcus , eg, Streptococcus agalactiae. According to yet another embodiment of the present disclosure, the Streptococcus is Group C Streptococcus , eg, Streptococcus equi, Streptococcus zooepidemicus, and Streptococcus nasus. According to yet another embodiment of the present disclosure, the Streptococcus is Group D Streptococcus (now reclassified as Enterococcus ), eg, Enterococcus faecalis , Enterococcus faecium , Enterococcus durans , and Enterococcus avium . According to other embodiments of the present disclosure, the Streptococcus is Group F Streptococcus , eg, Streptococcus anginosus (or Streptococcus milleri ), Streptococcus intermedius, and Streptococcus constellation. According to still other embodiments of the present disclosure, the Streptococcus is Group G Streptococcus , eg, Streptococcus nasus, Streptococcus canis, and Streptococcus zeta. According to yet other embodiments of the present disclosure, the Streptococcus is Group H Streptococcus , eg, Streptococcus sanguis. Other unclassified streptococci may also be suitable for use in the methods of the present disclosure, as long as the streptococcus can achieve the purpose of activating peripheral blood mononuclear cells to secrete inflammatory cytokines. Accordingly, streptococci suitable for use in the methods of the present disclosure include, but are not limited to, S. Streptococcus gordonii, Streptococcus intermedius, Streptococcus lentus, Streptococcus mutans, Streptococcus seals, Streptococcus pneumoniae, Streptococcus pyogenes, Streptococcus sialae, Streptococcus sanguis, Streptococcus suis, or Streptococcus zooepidemicus. In a preferred embodiment, the Streptococcus is type A Streptococcus, eg, Streptococcus pyogenes.

According to one embodiment of the present disclosure, the stimulator is a Gram-negative bacteria, including, but not limited to, Acinetobacter (eg, Acinetobacter baumannii ), Acinetobacter calcoaceticus ( Acinetobacter calocoaceticus ) , Acinetobacter lwoffi ); Bdellovibrio (eg, Bdellovibrio bacteriovorus ); Enterobacter (eg, Enterobacter cancerogenous) ), Enterobacter cloacae , Enterobacter cowanii , Enterobacter gergoviae , Enterobacter taylorae ); Escherichia (eg, Escherichia coli ) Haemophilus (eg, Haemophilus ducreyi , Haemophilus influenzae ); Helicobacter (eg, Helicobacter pylori ); Cree Klebsiella (eg, Klebsiella oxytoca , Klebsiella pneumoniae ); Legionella (eg, Legionella pneumophila ( Legionella pneumophila ); Moraxella (eg, Moraxella catarrhalis ); Neisseria (eg, Neisseria gonorrhoeae , Neisseria meningitidis ) ); Proteus (for example, Proteus mirabilis ); Pseudomonas (for example, Pseudomonas aeruginosa ), Pseudomonas oryzae ( Pseudomonas oryzihabitans ), Pseudomonas plecoglossicida ); Salmonella (eg, Salmonella bongori , Salmonella enteritidis , Salmonella typhi ); Serratia (eg, Serratia marcescens ); Shigella (eg, Shigella boydii ), Shigella dysenteriae, Shigella fukushima Shigella flexneri , Shigella sonnei ); Stenotrophomonas (eg, Stenotrophomonas maltophilia ). In a preferred embodiment, the Gram-negative bacteria is Escherichia coli.

In step (b), after the peripheral blood mononuclear cells are co-cultured with Streptococcus for a period of time (eg, 24 hours), the culture supernatant of the cells is collected to analyze that the cells are activated to secrete inflammatory cells hormone situation. The expression of inflammatory cytokines can be detected using a variety of assays well known to those skilled in the art, for example, an enzyme linked immunosorbent assay (ELISA) (e.g., multiplex assay), radioimmunoassay (RIA), immunofluorescence (IFA), Western blot (WB), immunoblotting (IB), immunoprecipitation (immunoprecipitation) , IP), or flow cytometry (flow cytometry) and other analytical methods. According to an operational embodiment of the present disclosure, ELISA is used to analyze the expression of inflammatory cytokines in the activated peripheral blood mononuclear cells.

Alternatively, peripheral blood mononuclear cells activated by Streptococcus to express the expression of inflammatory cytokines can be detected. The activated peripheral blood mononuclear cells can be collected, and the expression level of inflammatory cytokines can be detected by an analysis method for analyzing gene expression level. The analytical methods for analyzing gene expression levels are well known to those skilled in the art, including, but not limited to, genome-wide expression profiling using expressed sequence tag (EST) analysis Sequence analysis of gene expression (serial analysis of gene expression, SAGE); cDNA microarray (cDNA microarray); massively parallel signature sequencing (MPSS); RNA sequencing (RNA sequencing, RNA -seq); polymerase chain reaction (PCR) (for example, reverse transcription-PCR (RT-PCR), real-time RT-PCR or qRT-PCR ), digital polymerase chain reaction (digital-PCR or dPCR), touchdown PCR (touchdown PCR), nested PCR (nested PCR), multiplex PCR (multiplex PCR), recovery condition PCR (reconditioning PCR), etc.); two-dimensional coagulation Gel electrophoresis (two-dimensional gel electrophoresis, 2-D electrophoresis); tissue array (tissue array); immunohistochemistry (immunochemistry, IHC) staining and so on.

Next, after the peripheral blood mononuclear cells are activated, the plurality of drug candidates are tested using the activated peripheral blood mononuclear cells (step (c)). The specific step is to administer the drug candidates to be tested to the activated peripheral blood mononuclear cells, respectively, so that the activated peripheral blood mononuclear cells and the drug candidates to be tested are co-cultured for a period of time (for example, 16 to 72 hours, such as: 16, 18, 24, 36, 48, 60 or 72 hours; preferably, treatment for 24 hours) (step (c)). For the specific concentration of the candidate drug administered to the cells, reference may be made to the trough serum concentration under steady state recommended in the relevant pharmacokinetics section in the copy list of each candidate drug. For example, adalimumab (HUMIRA ^® ) use 4 μg/ml; golimumab (SIMPONI ^® ) use 0.5 μg/ml; guselkumab (TREMFYA ^® ) use 1.2 μg/ml; Monoclonal antibody (TALTZ ^® ) was used at 3.5 μg/ml; secukinumab (COSENTYX ^® ) at 34 μg/ml; and ustekinumab (STELARA ^® ) at 0.25 μg/ml.

The plurality of drug candidates may be current drugs for treating psoriasis, or potential drugs under development for treating psoriasis. According to certain embodiments of the present disclosure, the plurality of drug candidates comprises acitretin, afacept, allionol, apremilast, betamethasone, calcipotriene, calcipotriol, calcitriol Alcohol, Becloxone, Tar, Cyclosporine, Dicondol, Etanercept, Fluocinolone, Hydrocortisone, Infliximab, Ammethotrexate, Pimecrolimus, Tacrolimus, Tazarotene, Retinoic Acid, ALX-0761, BCD-085, Pestrolizumab, Bodalumab, CJM112, CNTO 6785, COVA322, Ixelizumab, LY3114062, MSB0010841, NI-1401, Pella Zizumab, rendolizumab, RG7624, secukinumab, vonezumab, blacumab, guselkumab, milizumab, risalizumab, tilta Xiximab, ustekinumab, adalimumab, certolizumab, etanercept, golimumab, infliximab, and/or lenacept. In addition, in a specific embodiment, the plurality of drug candidates comprises adalimumab, golimumab, guselkumab, ixelizumab, secukinumab, and ustekinumab anti.

In step (d), after the treatment of the specific candidate drug, the culture supernatant of the cells or the cells are collected respectively, and the expression of inflammatory cytokines caused by the influence of the drug on the cells is analyzed by the above analysis method Changes, particularly with respect to interferon-gamma, interleukin-13, and other inflammatory cytokines (eg, interleukin-6, interleukin-8, interleukin-12, or interleukin-17 etc.) changes in the amount of performance.

式(I)

Then, in step (e), a therapeutic drug is screened from the plurality of candidate drugs by using the changes in the expression levels of interferon-γ and interleukin-13 detected in step (d). The variation in the expression levels of interferon-γ and interleukin-13 refers to the expression levels of interferon-γ and interleukin-13 in step (b) (that is, before administration of a specific drug candidate), The relative rate of change with the expressed amounts of interferon-γ and interleukin-13 in step (d) (ie, after administration of a specific drug candidate). The relevant calculation formula is as follows, wherein formula (I) is a formula for calculating the rate of change of the expression of interferon-γ, and formula (II) is for calculating the ratio of the expression of interferon-γ to the expression of interleukin-13. The formula for the rate of change.

Formula (I)

According to a preferred embodiment of the present disclosure, in step (e), the therapeutic drug is selected from the plurality of candidate drugs with the highest inhibitory ability on the expression of interferon-γ (calculated according to formula (I) ) result), or select the drug with the highest inhibitory ability (or minimize the ratio) for the ratio of the expression of interferon-γ to the expression of interleukin-13 (according to the calculation result of formula (II)) medicine. According to an operating example, peripheral blood mononuclear cells of a specific psoriasis patient are treated with adalimumab and ustekinumab, wherein the administration of adalimumab can reduce the expression of interferon-γ in cells by approximately 9.4%, and administration of ustekinumab can reduce the expression of interferon-γ in cells by about 12.8%; in this example, ustekinumab was selected to treat the psoriasis patient. According to another working example, the peripheral blood mononuclear cells of a specific psoriasis patient are treated with Ixelizumab and ustekinumab, wherein the administration of Ixelizumab can increase the expression of interferon-gamma in the cells The ratio of expression to interleukin-13 increased by about 4.2%, while the administration of ustekinumab reduced the ratio of interferon-γ to interleukin-13 expression in cells by about 5.9%; In this example, ustekinumab was selected to treat the psoriasis patient.

Alternatively, those skilled in the art can screen therapeutic drugs based on the calculation results of the above formula (I). According to certain embodiments of the present disclosure, if a drug candidate reduces the amount of interferon-gamma expression by at least 1% (eg, a reduction of 1.0%, 1.1%, 1.2%, 1.3%, 1.4%, 1.5%, 1.6%) , 1.7%, 1.8%, 1.9%, 2.0%, 2.1%, 2.2%, 2.3%, 2.4%, 2.5%, 2.6%, 2.7%, 2.8%, 2.9%, 3.0%, 3.1%, 3.2%, 3.3 %, 3.4%, 3.5%, 3.6%, 3.7%, 3.8%, 3.9%, 4.0%, 4.1%, 4.2%, 4.3%, 4.4%, 4.5%, 4.6%, 4.7%, 4.8%, 4.9%, 5.0%, 5.1%, 5.2%, 5.3%, 5.4%, 5.5%, 5.6%, 5.7%, 5.8%, 5.9%, 6.0%, 6.1%, 6.2%, 6.3%, 6.4%, 6.5%, 6.6% , 6.7%, 6.8%, 6.9%, 7.0%, 7.1%, 7.2%, 7.3%, 7.4%, 7.5%, 7.6%, 7.7%, 7.8%, 7.9%, 8.0%, 8.1%, 8.2%, 8.3 %, 8.4%, 8.5%, 8.6%, 8.7%, 8.8%, 8.9%, 9.0%, 9.1%, 9.2%, 9.3%, 9.4%, 9.5%, 9.6%, 9.7%, 9.8%, 9.9%, 10.0%, 10.1%, 10.2%, 10.3%, 10.4%, 10.5%, 10.6%, 10.7%, 10.8%, 10.9%, 11.0%, 11.1%, 11.2%, 11.3%, 11.4%, 11.5%, 11.6% , 11.7%, 11.8%, 11.9%, 12.0%, 12.1%, 12.2%, 12.3%, 12.4%, 12.5%, 12.6%, 12.7%, 12.8%, 12.9%, 13.0%, 13.1%, 13.2%, 13.3 %, 13.4%, 13.5%, 13.6%, 13.7%, 13.8%, 13.9%, 14.0%, 14.1%, 14.2%, 14.3%, 14.4%, 14.5%, 14.6%, 14.7%, 14.8%, 14.9%, 15.0%, 15.1%, 15.2%, 15.3%, 15.4%, 15.5%, 15.6%, 15.7%, 15.8%, 15.9%, 16.0%, 16.1%, 16.2%, 16.3%, 16.4%, 16.5%, 16.6% , 16.7%, 16.8%, 16.9%, 17.0%, 17.1%, 17.2%, 17.3%, 17.4%, 17.5%, 17.6%, 17.7%, 17.8%, 17.9%, 18.0%, 18.1%, 18.2%, 18.3 %, 18. 4%, 18.5%, 18.6%, 18.7%, 18.8%, 18.9%, 19.0%, 19.1%, 19.2%, 19.3%, 19.4%, 19.5%, 19.6%, 19.7%, 19.8%, 19.9%, 20.0% , 25.0%, 30.0%, 35.0%, 40.0%, 45.0%, 50.0%, 55.0%, 60.0%, 65.0%, 70.0%, 75.0%, 80.0%, 85.0%, 90.0%, 95.0%, 100.0%, 110 %, 120%, 130%, 140%, 150%, 160%, 170%, 180%, 190%, 200%, 210%, 220%, 230%, 240%, 250%, 260%, 270%, 280%, 290%, or 300% expression of interferon-gamma), the candidate drug can be screened as a therapeutic drug. Preferably, if a drug candidate can reduce the expression of interferon-γ by about 1 to 20%, the drug candidate can be screened as a therapeutic drug. More preferably, if a drug candidate can reduce the expression of interferon-γ by about 1 to 10%, the drug candidate can be screened as a therapeutic drug. In certain embodiments, the drug candidate can reduce the amount of interferon-gamma expression by about 4.1%, 4.6%, 8.0%, 9.4%, 12.8%, 13.5%, 13.9%, 16.9%, respectively.

As can be imagined, those skilled in the art can also screen therapeutic drugs based on the calculation results of the above formula (II). According to certain alternative embodiments of the present disclosure, if a drug candidate reduces the ratio of interferon-gamma expression to interleukin-13 expression by at least 1% (eg, a reduction of 1.0%, 1.1%, 1.2%) , 1.3%, 1.4%, 1.5%, 1.6%, 1.7%, 1.8%, 1.9%, 2.0%, 2.1%, 2.2%, 2.3%, 2.4%, 2.5%, 2.6%, 2.7%, 2.8%, 2.9 %, 3.0%, 3.1%, 3.2%, 3.3%, 3.4%, 3.5%, 3.6%, 3.7%, 3.8%, 3.9%, 4.0%, 4.1%, 4.2%, 4.3%, 4.4%, 4.5%, 4.6%, 4.7%, 4.8%, 4.9%, 5.0%, 5.1%, 5.2%, 5.3%, 5.4%, 5.5%, 5.6%, 5.7%, 5.8%, 5.9%, 6.0%, 6.1%, 6.2% , 6.3%, 6.4%, 6.5%, 6.6%, 6.7%, 6.8%, 6.9%, 7.0%, 7.1%, 7.2%, 7.3%, 7.4%, 7.5%, 7.6%, 7.7%, 7.8%, 7.9 %, 8.0%, 8.1%, 8.2%, 8.3%, 8.4%, 8.5%, 8.6%, 8.7%, 8.8%, 8.9%, 9.0%, 9.1%, 9.2%, 9.3%, 9.4%, 9.5%, 9.6%, 9.7%, 9.8%, 9.9%, 10.0%, 10.1%, 10.2%, 10.3%, 10.4%, 10.5%, 10.6%, 10.7%, 10.8%, 10.9%, 11.0%, 11.1%, 11.2% , 11.3%, 11.4%, 11.5%, 11.6%, 11.7%, 11.8%, 11.9%, 12.0%, 12.1%, 12.2%, 12.3%, 12.4%, 12.5%, 12.6%, 12.7%, 12.8%, 12.9 %, 13.0%, 13.1%, 13.2%, 13.3%, 13.4%, 13.5%, 13.6%, 13.7%, 13.8%, 13.9%, 14.0%, 14.1%, 14.2%, 14.3%, 14.4%, 14.5%, 14.6%, 14.7%, 14.8%, 14.9%, 15.0%, 15.1%, 15.2%, 15.3%, 15.4%, 15.5%, 15.6%, 15.7%, 15.8%, 15.9%, 16.0%, 16.1%, 16.2% , 16.3%, 16.4%, 16.5%, 16.6%, 16.7%, 16.8%, 16.9%, 17.0%, 17.1%, 17.2%, 17.3%, 17.4%, 17.5%, 17.6%, 17.7%, 17.8%, 17.9 %, 18.0%, 18.1% , 18.2%, 18.3%, 18.4%, 18.5%, 18.6%, 18.7%, 18.8%, 18.9%, 19.0%, 19.1%, 19.2%, 19.3%, 19.4%, 19.5%, 19.6%, 19.7%, 19.8 %, 19.9%, 20.0%, 20.1%, 20.2%, 20.3%, 20.4%, 20.5%, 20.6%, 20.7%, 20.8%, 20.9%, 21.0%, 21.1%, 21.2%, 21.3%, 21.4%, 21.5%, 21.6%, 21.7%, 21.8%, 21.9%, 22.0%, 22.1%, 22.2%, 22.3%, 22.4%, 22.5%, 22.6%, 22.7%, 22.8%, 22.9%, 23.0%, 23.1% , 23.2%, 23.3%, 23.4%, 23.5%, 23.6%, 23.7%, 23.8%, 23.9%, 24.0%, 24.1%, 24.2%, 24.3%, 24.4%, 24.5%, 24.6%, 24.7%, 24.8 %, 24.9%, 25.0%, 25.1%, 25.2%, 25.3%, 25.4%, 25.5%, 25.6%, 25.7%, 25.8%, 25.9%, 26.0%, 26.1%, 26.2%, 26.3%, 26.4%, 26.5%, 26.6%, 26.7%, 26.8%, 26.9%, 27.0%, 27.1%, 27.2%, 27.3%, 27.4%, 27.5%, 27.6%, 27.7%, 27.8%, 27.9%, 28.0%, 28.1% , 28.2%, 28.3%, 28.4%, 28.5%, 28.6%, 28.7%, 28.8%, 28.9%, 29.0%, 29.1%, 29.2%, 29.3%, 29.4%, 29.5%, 29.6%, 29.7%, 29.8 %, 29.9%, 30.0%, 35.0%, 40.0%, 45.0%, 50.0%, 55.0%, 60.0%, 65.0%, 70.0%, 75.0%, 80.0%, 85.0%, 90.0%, 95.0%, 100.0%, 110%, 120%, 130%, 140%, 150%, 160%, 170%, 180%, 190%, 200%, 210%, 220%, 230%, 240%, 250%, 260%, 270% , 280%, 290%, or 300% of the expression of interferon-gamma to the expression of interleukin-13), the candidate drug can be screened as a therapeutic drug. Preferably, if a candidate drug can reduce the ratio of interferon-γ expression to interleukin-13 expression by about 1 to 30%, the candidate drug can be screened as a therapeutic drug. More preferably, if a candidate drug can reduce the ratio of interferon-γ expression to interleukin-13 expression by about 1 to 10%, the candidate drug can be screened as a therapeutic drug. In certain embodiments, the drug candidate can reduce the ratio of interferon-gamma expression to interleukin-13 expression by about 1.1%, 4.3%, 4.6%, 5.9%, 9.2%, and 27.1%, respectively.

2. treatment method

Another aspect of the present disclosure pertains to a method for treating psoriasis in an individual comprising: (1) Screen a therapeutic drug from a plurality of candidate drugs by the following steps, wherein the therapeutic drug is used to treat the psoriasis of the individual, including: (1a) Obtaining an isolated peripheral blood mononuclear cell from the individual; (1b) activate the peripheral blood mononuclear cells of step (1a) with a stimulant, so that the cells produce interferon-γ and interleukin-13; (1c) respectively administering the plurality of candidate drugs to the activated peripheral blood mononuclear cells in step (1b); (1d) respectively detecting the expression levels of the interferon-γ and the interleukin-13 in the peripheral blood mononuclear cells of step (1c); and (1e) Based on the expression levels of the interferon-γ and the interleukin-13 detected in step (1d), the therapeutic drug is selected from the plurality of candidate drugs, wherein the therapeutic drug is the interferon-γ The expression of gamma has the highest inhibitory ability, or the ratio of the expression of interferon-gamma to the expression of interleukin-13 has the highest inhibitory ability; and (2) Administer an effective amount of the therapeutic drug of step (1e) to the individual.

In the treatment method of the present disclosure, the steps of screening the therapeutic drugs (that is, the steps described in step (1), including steps (1a) to (1e)) are the same as the screening methods of the present disclosure. For the sake of brevity, here No longer.

Step (2) is to administer the therapeutic drug screened in step (1) to the individual, which can be administered to the individual by oral, intravenous, intradermal, subcutaneous, transdermal, topical application, or intramuscular routes. The pharmaceutical compositions of the present disclosure are administered. According to certain embodiments of the present disclosure, an effective amount of a therapeutic drug is administered to the individual via intravenous injection to achieve the therapeutic purpose.

The subject is a mammal including humans, rats, mice, guinea pigs, monkeys, pigs, goats, cows, horses, dogs, cats, birds and chickens. Preferably, the individual is a human.

Individuals treated with the treatment methods of the present disclosure can be assessed for treatment efficacy by the Psoriasis Area Severity Index (PASI). The evaluation method of the PASI covers the evaluation of the psoriasis area (Area) and the psoriasis severity (Severity), as shown in Table 1 and Table 2, and after evaluating the psoriasis area and psoriasis severity, the value obtained is by the following formula To calculate PASI: PASI=0.1×(E _h ＋I _h ＋D _h )×A _h ＋0.3×(E _t ＋I _t ＋D _t )×A _t ＋0.2×(E _u ＋I _u ＋D _u )×A _u ＋0 .4×(E _l +I _l +D _l )×A _l ; Parts: head (h), trunk (t), upper limbs (u), lower limbs (l). Table 1 Psoriasis area (Area): combined total area % of body surface Coverage 0% <10% 10-29% 30-49% 50-69% 70-89% 90-100% Score Skin Site 0 1 2 3 4 5 6 Location: head (h), trunk (t), upper extremity (u), lower extremity (l) Table 2 Psoriasis severity (Severity): Severity none slight Moderate severe most serious Fraction 0 1 2 3 4 redness (E) none slightly pink Pink red dark red/purpuric Lesion thickness (I) none slight thickening Moderate thickening with rounded or sloping edges Significantly thickened and bordered Very noticeable thickening with very hard and noticeable edges Desquamation condition (D) none Slight scaling Extensive desquamation and most of the lesions partially covered by desquamation Extensive desquamation and almost all lesions covered by rough surface All lesions covered with very large and thick desquamation, very rough surface

Individuals treated by the methods of the present disclosure may find that the PASI is significantly improved before and after treatment. For example, the PASI after treatment can be improved by at least 3 points or more compared to the PASI before treatment, such as 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24 or 25 points.

Several experimental examples are provided below to illustrate certain aspects of the present invention, so as to facilitate the practice of the present invention by those skilled in the art to which the present invention pertains, and these experimental examples should not be regarded as limiting the scope of the present invention. It is believed that those skilled in the art, after reading the description presented herein, can fully utilize and practice the present invention without undue interpretation. All publications cited herein are considered part of this specification in their entirety.

Example

Materials and Methods

1. Patient and clinical samples

This study employed a group of Taiwanese patients with psoriasis, and all participants had obtained written informed consent before completing sample collection.

2. Cell isolation and culture

16 ml of peripheral blood (whole blood) from patients with psoriasis was collected into a cell preparation tube (CPT). Next, the blood was centrifuged at 1700×g for 15 minutes, and peripheral blood mononuclear cells were then aspirated with a 3 ml pipette. Afterwards, cells were washed with phosphate buffered saline (PBS) and cultured in RPMI 1640 medium supplemented with 10% fetal bovine serum (FBS) and 1% antibiotics. Cells were grown at 37°C in a 5% carbon dioxide incubator.

Regarding the activation of peripheral blood mononuclear cells and testing drug candidates, the concentration of peripheral blood mononuclear cells and the bacterial solution of Streptococcus pyogenes (Streptococcus pyogenes) McFarland 2 (McFarland 2) solution and different biological agents After 24 hours of drug candidate co-culture, cell supernatants were collected for analysis of inflammatory hormones (including interferon-γ, interleukin-13, and other inflammatory cytokines (eg, interleukin-6, interleukin-8, Interleukin-12, or interleukin-17, etc.)) concentration. The concentration of the candidate biologics tested in this study was based on the trough serum concentration at steady state as disclosed in the pharmacokinetics section of each biologics copy. The specific concentrations used are as follows: Adalimumab (HUMIRA ^® ) uses 4 μg/ml; Golimumab (SIMPONI ^® ) uses 0.5 μg/ml; Guselkumab (TREMFYA ^® ) uses 1.2 μg/ml; Use 3.5 micrograms/ml for beizumab (TALTZ ^® ); 34 micrograms/ml for secukinumab (COSENTYX ^® ); and 0.25 micrograms/ml for ustekinumab (STELARA ^® ).

3. Statistical Analysis

All quantitative experimental data are presented as mean ± standard deviation (Mean ± SD) unless otherwise stated. Spearman's rank correlation coefficient was used to analyze: (1) the change ratio or difference of PASI obtained before and after treatment, and (2) the obtained peripheral blood mononuclear cells of psoriasis patients before and after the administration of the candidate drug either (a) the ratio or difference in the ratio of inflammatory cytokines (eg, interferon-gamma), or (b) the ratio of changes in the ratio of inflammatory cytokines (eg, interferon-gamma/interleukin-13) or the difference, the correlation between the two. Two-tailed Student's t-test was used to compare different samples, and P < 0.05 was considered statistically significant; *: P < 0.05; **: P < 0.01.

Example 1 before and after treatment PASI Relationship between changes and changes in inflammatory cytokines

Because different patients with psoriasis respond differently to different biologics, it is critical to develop a method of evaluating medication for each patient with psoriasis to assess the appropriate therapy for that patient. This study therefore developed the use of the expression change of inflammatory cytokines or the ratio of inflammatory cytokines produced by peripheral blood mononuclear cells before and after administration as a medication index to assess whether the patient is suitable for a specific therapy. In this example, the relationship between changes in PASI and changes (ie, ratios or differences) in the expression of inflammatory cytokines in different patients before and after treatment with a specific biologic was analyzed.

Example 1.1 Interferon -γ

The relationship between the changes of PASI before and after treatment and the changes of interferon-γ before and after administration and related statistical analysis are shown in Tables 3 to 4. Among them, the change of PASI before and after treatment refers to the change of PASI value of individual patients before and after receiving individual biological agents; and the change of interferon-γ before and after administration refers to the measurement of peripheral blood of the patient. Mononuclear cells were tested in vitro for changes in interferon-gamma before and after drug administration for individual biologics using the methods of drug screening described in this disclosure. When calculating the change of PASI, the calculation method of the difference value is: after treatment-before treatment; and the calculation method of the change ratio is: [(after treatment-before treatment)/before treatment]×100%. When calculating the change amount of interferon-γ, the calculation method of the difference is: after administration-before administration; and the calculation method of the change ratio is: [(after administration-before administration)/before administration]×100 %.

Table 3 Changes of PASI and interferon-γ before and after treatment patient Types of biologics used PASI (before treatment) PASI (after treatment) PASI change ratio PASI difference A ^HUMIRA® 3.2 2.6 -18.8% -0.6 A ^STELARA® 3.2 0 -100.0% -3.2 B ^HUMIRA® 15 16 6.7% 1 B ^STELARA® 15 0.2 -98.7% -14.8 C ^HUMIRA® 13 0.5 -96.2% -12.5 C ^STELARA® 13 12 -7.7% -1 D ^HUMIRA® 7.5 15 100.0% 7.5 D ^STELARA® 7.5 0.8 -89.3% -6.7 E ^HUMIRA® 25 0.4 -98.4% -24.6 E TREMFYA ^® 8 4.3 -46.3% -3.7 E ^COSENTYX® 8 1.2 -85.0% -6.8 patient Types of biologics used Interferon-gamma (pre-dose) (pg/ml) Interferon-gamma (post-dose) (pg/ml) Interferon-gamma change ratio Interferon-gamma difference (pg/ml) A ^HUMIRA® 13.72 12.43 -9.4% -1.29 A ^STELARA® 13.72 11.97 -12.8% -1.75 B ^HUMIRA® 74.46 88.28 18.6% 13.82 B ^STELARA® 74.46 71.02 -4.6% -3.44 C ^HUMIRA® 22.9 19.71 -13.9% -3.19 C ^STELARA® 22.9 21.97 -4.1% -0.93 D ^HUMIRA® 200.39 222.11 10.8% 21.72 D ^STELARA® 200.39 201.66 0.6% 1.27 E ^HUMIRA® 92.6 76.99 -16.9% -15.61 E TREMFYA ^® 92.6 85.17 -8.0% -7.43 E ^COSENTYX® 92.6 80.12 -13.5% -12.48

Table 4 Relevant Statistical Results of Table 3 PASI change ratio to interferon -gamma change ratio PASI change ratio versus interferon -gamma difference Spearman coefficient r r 0.6364 -0.6182 95% confidence interval 0.03841 to 0.8987 -0.8928 to -0.008442 P value P value (two-tailed test) 0.0402 0.0478 P value results * * Is it significant? (α = 0.05) Significant Significant PASI difference to interferon -gamma change ratio PASI difference vs. interferon -gamma difference Spearman coefficient r r -0.6273 0.7818 95% confidence interval -0.8958 to -0.02329 0.3243 to 0.9429 P value P value (two-tailed test) 0.044 0.0064 P value results * ** Is it significant? (α = 0.05) Significant Significant

Based on the results in Table 3 above, it can be seen that, for a specific psoriasis patient, administration of different biological agents for treatment will cause the change ratio or difference of its PASI to decrease to varying degrees (that is, the patient's psoriasis condition is obtained to varying degrees. At the same time, it was also found that the change ratio or difference of the amount of interferon-γ expression would decrease in different degrees. For example, for patient A, compared to before administration of HUMIRA ^® or STELARA ^® , the rate of change in PASI decreased by 18.8% and 100.0%, respectively, and the difference in PASI decreased after treatment. 0.6 and 3.2; at the same time, the rate of change in the amount of interferon-γ expression decreased by 9.4% and 12.8%, respectively, and the difference in the amount of interferon-γ expression decreased by 1.29 and 1.75 pg/ml, respectively, and so on. Accordingly, the present example further analyzes the relationship between PASI and interferon-γ changes before and after treatment by statistical analysis, and is summarized in Table 4. Based on the results in Table 4 above, it can be seen that the reduction ratio of PASI before and after treatment with different biological agents is significantly related to the reduction ratio or the difference of the patient's interferon-γ, and the difference of PASI reduction is related to the patient's interferon-gamma. Both the ratio of declines or the difference in declines in γ were significantly correlated. Accordingly, the change in the expression of interferon-γ before and after treatment (whether it is the decreasing ratio or the decreasing difference) can be used as a medication index for psoriasis patients.

Example 1.2 between white pigment -6

The relationship between the changes of PASI before and after treatment and the changes of interleukin-6 before and after administration and related statistical analysis are shown in Tables 5 to 6. Among them, the change in PASI before and after treatment refers to the change in PASI values recorded by individual patients before and after receiving individual biological agents; and the change in interleukin-6 before and after administration refers to taking the peripheral Blood monocytes were tested in vitro for changes in interleukin-6 before and after drug administration for individual biologics using the methods of drug screening described in this disclosure. When calculating the change of PASI, the calculation method of the difference value is: after treatment-before treatment; and the calculation method of the change ratio is: [(after treatment-before treatment)/before treatment]×100%. When calculating the change amount of interleukin-6, the calculation method of the difference is: after administration-before administration; and the calculation method of the change ratio is: [(after administration-before administration)/before administration]× 100%.

Table 5 Changes of PASI and interleukin-6 before and after treatment patient Types of biologics used PASI (before treatment) PASI (after treatment) PASI change ratio PASI difference A ^HUMIRA® 3.2 2.6 -18.8% -0.6 A ^STELARA® 3.2 0 -100.0% -3.2 B ^HUMIRA® 15 16 6.7% 1 B ^STELARA® 15 0.2 -98.7% -14.8 C ^HUMIRA® 13 0.5 -96.2% -12.5 C ^STELARA® 13 12 -7.7% -1 D ^HUMIRA® 7.5 15 100.0% 7.5 D ^STELARA® 7.5 0.8 -89.3% -6.7 E ^HUMIRA® 25 0.4 -98.4% -24.6 E TREMFYA ^® 8 4.3 -46.3% -3.7 E ^COSENTYX® 8 1.2 -85.0% -6.8 patient Types of biologics used Interleukin-6 (pre-dose) (pg/ml) Interleukin-6 (after administration) (pg/ml) Interleukin-6 Alteration Ratio Interleukin-6 Difference (Pg/mL) A ^HUMIRA® 269.08 253.7 -5.7% -15.38 A ^STELARA® 269.08 233.74 -13.1% -35.34 B ^HUMIRA® 1992.34 2134.34 7.1% 142 B ^STELARA® 1992.34 2079.52 4.4% 87.18 C ^HUMIRA® 468.01 425.03 -9.2% -42.98 C ^STELARA® 468.01 455.88 -2.6% -12.13 D ^HUMIRA® 8985.92 9421.22 4.8% 435.3 D ^STELARA® 8985.92 8574.26 -4.6% -411.66 E ^HUMIRA® 12915.64 12207.14 -5.5% -708.5 E TREMFYA ^® 12915.64 14041.18 8.7% 1125.54 E ^COSENTYX® 12915.64 12819.59 -0.7% -96.05

Table 6 Table 5 Relevant Statistical Results PASI change ratio to interleukin - 6 change ratio PASI change ratio versus interleukin - 6 difference Spearman coefficient r r 0.5636 -0.5455 95% confidence interval -0.07532 to 0.8745 -0.8681 to 0.1014 P value P value (two-tailed test) 0.0761 0.0876 P value results not significant not significant Is it significant? (α = 0.05) not significant not significant PASI difference to interleukin - 6 change ratio PASI difference versus interleukin - 6 difference Spearman coefficient r r -0.2727 0.6 95% confidence interval -0.7588 to 0.4085 -0.02046 to 0.8868 P value P value (two-tailed test) 0.4181 0.0562 P value results not significant not significant Is it significant? (α = 0.05) not significant not significant

Based on the results in Table 5 above, it can be seen that for a specific psoriasis patient, the change ratio or difference of PASI will be reduced to varying degrees when different biological agents are administered for treatment. However, the change in the expression of interleukin-6 The ratio or difference produces an up or down change. For example, for patient A, compared to before administration of HUMIRA ^® or STELARA ^® , the rate of change in PASI decreased by 18.8% and 100.0%, respectively, and the difference in PASI decreased after treatment. 0.6 and 3.2; at the same time, the change rate of its interleukin-6 expression decreased by 5.7% and 13.1%, respectively, and the difference of its interleukin-6 expression decreased by 15.38 and 35.34 pg/ml, respectively, and so on. . Accordingly, in order to understand the relationship between the changes of PASI and interleukin-6, statistical analysis was further performed in this example, and the relevant results are provided in Table 6. Based on the results in Table 6 above, it can be seen that the reduction ratio of PASI before and after treatment with different biological agents is not significantly related to the reduction ratio or the difference of the patient's interleukin-6, and the difference of the PASI reduction is related to the patient's interleukin-6. There was also no significant correlation between the rate of decline or the difference in decline in leukin-6.

Example 1.3 between white pigment -8

The relationship between the changes of PASI before and after treatment and the changes of interleukin-8 before and after administration and related statistical analysis are shown in Tables 7 to 8. Among them, the change in PASI before and after treatment refers to the change in PASI values recorded by individual patients before and after receiving individual biological agents; and the change in interleukin-8 before and after administration refers to taking the peripheral Blood monocytes were tested in vitro for changes in interleukin-8 before and after drug administration for individual biologics using the methods of drug screening described in this disclosure. When calculating the change of PASI, the calculation method of the difference value is: after treatment-before treatment; and the calculation method of the change ratio is: [(after treatment-before treatment)/before treatment]×100%. When calculating the change amount of interleukin-8, the calculation method of the difference is: after administration-before administration; and the calculation method of the change ratio is: [(after administration-before administration)/before administration]× 100%.

Table 7 Changes of PASI and interleukin-8 before and after treatment patient Types of biologics used PASI (before treatment) PASI (after treatment) PASI change ratio PASI difference A ^HUMIRA® 3.2 2.6 -18.8% -0.6 A ^STELARA® 3.2 0 -100.0% -3.2 B ^HUMIRA® 15 16 6.7% 1 B ^STELARA® 15 0.2 -98.7% -14.8 C ^HUMIRA® 13 0.5 -96.2% -12.5 C ^STELARA® 13 12 -7.7% -1 D ^HUMIRA® 7.5 15 100.0% 7.5 D ^STELARA® 7.5 0.8 -89.3% -6.7 E ^HUMIRA® 25 0.4 -98.4% -24.6 E TREMFYA ^® 8 4.3 -46.3% -3.7 E ^COSENTYX® 8 1.2 -85.0% -6.8 patient Types of biologics used Interleukin-8 (pre-dose) (pg/ml) Interleukin-8 (post-administration) (pg/ml) Interleukin-8 Altered Ratio Interleukin-8 Difference (Pg/mL) A ^HUMIRA® 4002.25 3212.29 -19.7% -789.96 A ^STELARA® 4002.25 3622.79 -9.5% -379.46 B ^HUMIRA® 27693.97 431959.9 1459.8% 404265.93 B ^STELARA® 27693.97 36870.27 33.1% 9176.3 C ^HUMIRA® 6952.5 7980.57 14.8% 1028.07 C ^STELARA® 6952.5 8082.76 16.3% 1130.26 D ^HUMIRA® 9204.67 11061.83 20.2% 1857.16 D ^STELARA® 9204.67 8091.09 -12.1% -1113.58 E ^HUMIRA® 13225.99 12401.19 -6.2% -824.8 E TREMFYA ^® 13225.99 13132.35 -0.7% -93.64 E ^COSENTYX® 13225.99 11597.1 -12.3% -1628.89

Relevant statistical results of Table 8 and Table 7 PASI change ratio to interleukin - 8 ratio PASI change ratio versus interleukin - 8 difference Spearman coefficient r r 0.2636 -0.3364 95% confidence interval -0.4166 to 0.7546 -0.7870 to 0.3484 P value P value (two-tailed test) 0.4348 0.3132 P value results not significant not significant Is it significant? (α = 0.05) not significant not significant PASI difference to interleukin - 8 change ratio PASI Difference vs Interleukin - 8 Difference Spearman coefficient r r -0.1727 0.3818 95% confidence interval -0.7104 to 0.4923 -0.3017 to 0.8061 P value P value (two-tailed test) 0.6147 0.2483 P value results not significant not significant Is it significant? (α = 0.05) not significant not significant

Based on the results in Table 7 above, it can be seen that, for a specific psoriasis patient, the ratio or difference of PASI changes will be reduced to varying degrees when different biological agents are administered for treatment. The ratio or difference produces an up or down change. For example, for patient A, compared to before administration of HUMIRA ^® or STELARA ^® , the rate of change in PASI decreased by 18.8% and 100.0%, respectively, and the difference in PASI decreased after treatment. 0.6 and 3.2; at the same time, the change rate of its interleukin-8 expression decreased by 19.7% and 9.5%, respectively, and the difference of its interleukin-8 expression decreased by 789.96 and 379.46 pg/ml, and so on. . Accordingly, in order to understand the relationship between the changes of PASI and interleukin-8, statistical analysis was further performed in this example, and the relevant results are provided in Table 8. Based on the results in Table 8 above, it can be seen that the reduction ratio of PASI before and after treatment with different biological agents is not significantly related to the reduction ratio or the difference of the patient's interleukin-8, and the difference of PASI reduction is related to the patient's interleukin-8. There was also no significant correlation between the rate of decline in protein-8 or the difference in decline.

Example 1.4 between white pigment -12

The relationship between the changes of PASI before and after treatment and the changes of interleukin-12 before and after administration and related statistical analysis are shown in Table 9 to Table 10. Among them, the change in PASI before and after treatment refers to the change in PASI values recorded by individual patients before and after receiving individual biological agents; and the change in interleukin-12 before and after administration refers to the patient Blood mononuclear cells were tested in vitro for changes in interleukin-12 before and after drug administration for individual biologics using the methods of drug screening described in this disclosure. When calculating the change of PASI, the calculation method of the difference value is: after treatment-before treatment; and the calculation method of the change ratio is: [(after treatment-before treatment)/before treatment]×100%. When calculating the change amount of interleukin-12, the calculation method of the difference is: after administration-before administration; and the calculation method of the change ratio is: [(after administration-before administration)/before administration]× 100%.

Table 9 Changes of PASI and interleukin-12 before and after treatment patient Types of biologics used PASI (before treatment) PASI (after treatment) PASI change ratio PASI difference A ^HUMIRA® 3.2 2.6 -18.8% -0.6 A ^STELARA® 3.2 0 -100.0% -3.2 B ^HUMIRA® 15 16 6.7% 1 B ^STELARA® 15 0.2 -98.7% -14.8 C ^HUMIRA® 13 0.5 -96.2% -12.5 C ^STELARA® 13 12 -7.7% -1 D ^HUMIRA® 7.5 15 100.0% 7.5 D ^STELARA® 7.5 0.8 -89.3% -6.7 E ^HUMIRA® 25 0.4 -98.4% -24.6 E TREMFYA ^® 8 4.3 -46.3% -3.7 E ^COSENTYX® 8 1.2 -85.0% -6.8 patient Types of biologics used Interleukin-12 (before administration) (pg/ml) Interleukin-12 (after administration) (pg/ml) Interleukin-12 Altered Ratio Interleukin-12 Difference (Pg/mL) A ^HUMIRA® 1.86 1.86 0.0% 0 A ^STELARA® 1.86 1.86 0.0% 0 B ^HUMIRA® 8.56 11.13 30.0% 2.57 B ^STELARA® 8.56 5.67 -33.8% -2.89 C ^HUMIRA® 3.05 3.63 19.0% 0.58 C ^STELARA® 3.05 3.34 9.5% 0.29 D ^HUMIRA® 12 11.55 -3.7% -0.45 D ^STELARA® 12 11.89 -0.9% -0.11 E ^HUMIRA® 9.28 7.05 -24.0% -2.23 E TREMFYA ^® 9.28 7.97 -14.1% -1.31 E ^COSENTYX® 9.28 9.7 4.5% 0.42

Table 10 Relevant Statistical Results of Table 9 PASI Alteration Ratio to Interleukin - 12 Ratio PASI change ratio versus interleukin - 12 difference Spearman coefficient r r 0.3599 -0.3326 95% confidence interval -0.3246 to 0.7970 -0.7854 to 0.3521 P value P value (two-tailed test) 0.2757 0.3082 P value results not significant not significant Is it significant? (α = 0.05) not significant not significant PASI difference to interleukin - 12 change ratio PASI difference versus interleukin - 12 difference Spearman coefficient r r -0.3918 0.3554 95% confidence interval -0.8102 to 0.2910 -0.3293 to 0.7951 P value P value (two-tailed test) 0.2271 0.2814 P value results not significant not significant Is it significant? (α = 0.05) not significant not significant

Based on the results in Table 9 above, it can be seen that, for a specific psoriasis patient, administration of different biological agents for treatment, the change ratio or difference of PASI will decrease to varying degrees, however, the change in the expression of interleukin-12 The ratio or difference produces an up or down change. For example, for patient B, compared with before the administration of HUMIRA ^® or STELARA ^® , the rate of change in PASI increased by 6.7% and decreased by 98.7%, respectively, and the difference in PASI was respectively It increased by 1 and decreased by 14.8; at the same time, the change rate of its interleukin-12 expression increased by 30.0% and decreased by 33.8%, and the difference of its interleukin-12 expression increased by 2.57 and decreased by 2.89 pg/ml, respectively. , and so on. Accordingly, in order to understand the relationship between PASI and interleukin-12 variation, statistical analysis was further performed in this example, and the relevant results are provided in Table 10. Based on the results in Table 10 above, it can be seen that the reduction ratio of PASI before and after treatment with different biological agents has no significant correlation with the reduction ratio or the difference of the patient's interleukin-12, and the difference of PASI reduction is related to the patient's interleukin-12. There was also no significant correlation between the rate of decline or the difference in decline in leukin-12.

Example 1.5 between white pigment -17

The relationship between the changes of PASI before and after treatment and the changes of interleukin-17 before and after administration and related statistical analysis are shown in Table 11 to Table 12. Among them, the change in PASI before and after treatment refers to the change in PASI values recorded by individual patients before and after receiving individual biological agents; and the change in interleukin-17 before and after administration refers to the patient Individual biologics were tested in vitro for changes in interleukin-17 before and after drug administration on blood mononuclear cells using the methods of drug screening described in this disclosure. When calculating the change of PASI, the calculation method of the difference value is: after treatment-before treatment; and the calculation method of the change ratio is: [(after treatment-before treatment)/before treatment]×100%. When calculating the change amount of interleukin-17, the calculation method of the difference is: after administration-before administration; and the calculation method of the change ratio is: [(after administration-before administration)/before administration]× 100%.

Table 11 Changes of PASI and interleukin-17 before and after treatment patient Types of biologics used PASI (before treatment) PASI (after treatment) PASI change ratio PASI difference A ^HUMIRA® 3.2 2.6 -18.8% -0.6 A ^STELARA® 3.2 0 -100.0% -3.2 B ^HUMIRA® 15 16 6.7% 1 B ^STELARA® 15 0.2 -98.7% -14.8 C ^HUMIRA® 13 0.5 -96.2% -12.5 C ^STELARA® 13 12 -7.7% -1 D ^HUMIRA® 7.5 15 100.0% 7.5 D ^STELARA® 7.5 0.8 -89.3% -6.7 E ^HUMIRA® 25 0.4 -98.4% -24.6 E TREMFYA ^® 8 4.3 -46.3% -3.7 E ^COSENTYX® 8 1.2 -85.0% -6.8 patient Types of biologics used Interleukin-17 (before administration) (pg/ml) Interleukin-17 (after administration) (pg/ml) Interleukin-17 Altered Ratio Interleukin-17 Difference (Pg/mL) A ^HUMIRA® 69.02 65.58 -5.0% -3.44 A ^STELARA® 69.02 68.26 -1.1% -0.76 B ^HUMIRA® 109.09 78.53 -28.0% -30.56 B ^STELARA® 109.09 107.2 -1.7% -1.89 C ^HUMIRA® 109.76 103.76 -5.5% -6 C ^STELARA® 109.76 103.76 -5.5% -6 D ^HUMIRA® 179.24 185.8 3.7% 6.56 D ^STELARA® 179.24 193.12 7.7% 13.88 E ^HUMIRA® 85.99 67.64 -21.3% -18.35 E TREMFYA ^® 85.99 82.1 -4.5% -3.89 E ^COSENTYX® 85.99 82.1 -4.5% -3.89

Table 12 Relevant Statistical Results of Table 11 PASI change ratio to interleukin - 17 change ratio PASI change ratio versus interleukin - 17 difference Spearman coefficient r r -0.1827 0.1553 95% confidence interval -0.7155 to 0.4845 -0.5058 to 0.7014 P value P value (two-tailed test) 0.5792 0.6472 P value results not significant not significant Is it significant? (α = 0.05) not significant not significant PASI difference to interleukin - 17 change ratio PASI difference versus interleukin - 17 difference Spearman coefficient r r -0.07763 0.1507 95% confidence interval -0.6592 to 0.5621 -0.5093 to 0.6991 P value P value (two-tailed test) 0.8119 0.657 P value results not significant not significant Is it significant? (α = 0.05) not significant not significant

Based on the results in Table 11 above, it can be seen that, for a specific psoriasis patient, the ratio or difference of PASI changes will be reduced to varying degrees when different biological agents are administered for treatment. However, the change in the expression of interleukin-17 The ratio or difference produces an up or down change. For example, for patient A, compared to before administration of HUMIRA ^® or STELARA ^® , the rate of change in PASI decreased by 18.8% and 100.0%, respectively, and the difference in PASI decreased after treatment. 0.6 and 3.2; at the same time, the change rate of its interleukin-17 expression decreased by 5.0% and 1.1%, and the difference of its interleukin-17 expression decreased by 3.44 and 0.76 pg/ml, respectively, and so on. . Accordingly, in order to understand the relationship between PASI and interleukin-17 variation, statistical analysis was further performed in this example, and the relevant results are provided in Table 12. Based on the results in Table 12 above, it can be seen that the reduction ratio of PASI before and after treatment with different biological agents is not significantly related to the reduction ratio or the difference of the patient's interleukin-17, and the difference of PASI reduction is related to the patient's interleukin-17. There was also no significant correlation between the rate of decline or the difference in decline in leukin-17.

Summarizing the above results, this example proves that interferon-γ will produce a change in the expression of decreased expression (whether it is the reduction ratio or the difference) before and after the treatment of different biological agents, so the decrease in the expression of interferon-γ can be regarded as psoriasis. patient medication indicators.

Example 2 before and after treatment PASI Changes and Interferon -γ/ between white pigment -13 The relationship between the change in the ratio of

In this example, the relationship between the change in PASI and the expressive change in the ratio of inflammatory cytokines (ie, change ratio or difference) in different patients before and after treatment with a specific biological agent was analyzed. Table 13 to Table 14 show the relationship between the change in PASI before and after treatment and the change in the ratio of interferon-γ/interleukin-13 before and after administration and the related statistical analysis. Among them, the change in PASI before and after treatment refers to the change in the PASI value of individual patients before and after receiving individual biological agents; and the change in the ratio of interferon-γ/interleukin-13 before and after administration is Refers to the interferon-γ/interleukin-13 before and after drug administration obtained by taking the peripheral blood mononuclear cells of the patient and testing individual biologics in vitro by the method of drug screening described in this disclosure The amount of change in the ratio. Regarding the part of calculating the change in PASI, the calculation method of the difference is: after treatment-before treatment; and the calculation method of the change ratio is: [(after treatment-before treatment)/before treatment]×100%. And in the part of the change in the ratio of interferon-γ/interleukin-13, the calculation method of the difference is: obtained after administration (ratio of interferon-γ/interleukin-13) - before administration The obtained (interferon-γ/interleukin-13 ratio); and the calculation method of the change ratio is shown in the above formula (II).

Table 13 Changes in the ratio of PASI to interferon-γ/interleukin-13 before and after treatment patient Types of biologics used PASI (before treatment) PASI (after treatment) PASI change ratio PASI difference A ^TALTZ® 15 1.2 -92.0% -13.8 A ^STELARA® 16 0 -100.0% -16 B ^HUMIRA® 15 16 6.7% 1 B ^STELARA® 15 0.2 -98.7% -14.8 C ^HUMIRA® 13 0.5 -96.2% -12.5 C ^STELARA® 13 12 -7.7% -1 D ^HUMIRA® 7.5 15 100.0% 7.5 D ^STELARA® 7.5 0.8 -89.3% -6.7 E ^HUMIRA® 25 12 -52.0% -13 E TREMFYA ^® 8 4.3 -46.3% -3.7 E ^COSENTYX® 8 1.2 -85.0% -6.8 patient Types of biologics used Interferon-gamma/interleukin-13 ratio (before administration) Interferon-gamma/interleukin-13 ratio (after administration) Changes in the ratio of interferon-gamma/interleukin-13 Interferon-gamma/interleukin-13 ratio difference A ^TALTZ® 127.3 132.63 4.2% 5.33 A ^STELARA® 127.3 119.85 -5.9% -7.45 B ^HUMIRA® 60.05 108.99 81.5% 48.94 B ^STELARA® 60.05 57.27 -4.6% -2.78 C ^HUMIRA® 45.8 33.4 -27.1% -12.4 C ^STELARA® 45.8 43.82 -4.3% -1.98 D ^HUMIRA® 38.6 51.53 33.5% 12.93 D ^STELARA® 38.6 38.19 -1.1% -0.41 E ^HUMIRA® 141.95 151.67 6.8% 9.72 E TREMFYA ^® 141.95 155.73 9.7% 13.78 E ^COSENTYX® 141.95 128.9 -9.2% -13.05

Table 14 Relevant Statistical Results of Table 13 The ratio of change in PASI to the ratio of interferon -γ/ interleukin - 13 The difference between the ratio of PASI change and the ratio of interferon -γ/ interleukin - 13 Spearman coefficient r r 0.7364 -0.6818 95% confidence interval 0.2250 to 0.9297 -0.9131 to -0.1183 P value P value (two-tailed test) 0.0128 0.0251 P value results Significant Significant Is it significant? (α = 0.05) Significant Significant Change ratio of PASI difference to interferon -γ/ interleukin - 13 ratio The difference between PASI difference and interferon -γ/ interleukin - 13 ratio Spearman coefficient r r -0.5818 0.5455 95% confidence interval -0.8807 to 0.04837 -0.1014 to 0.8681 P value P value (two-tailed test) 0.0656 0.0876 P value results not significant not significant Is it significant? (α = 0.05) not significant not significant

Based on the results in Table 13 above, it can be seen that for a specific psoriasis patient, administration of different biological agents for treatment will cause the change ratio or difference of PASI to decrease to varying degrees. At the same time, it is also found that interferon-γ The change ratio or difference of the ratio of /IL-13 will decrease in different degrees. For example, in patient C, compared to before the administration of HUMIRA ^® or STELARA ^® , the rate of change in PASI decreased by 96.2% and 7.7%, respectively, and the difference in PASI decreased, respectively. 12.5 and 1; at the same time, the change ratio of its interferon-gamma/interleukin-13 ratio decreased by 27.1% and 4.3%, respectively, and the difference of its interferon-gamma/interleukin-13 ratio decreased by 12.4 and 1.98, and so on. Accordingly, the present example further analyzes the relationship between PASI and the ratio of interferon-γ/interleukin-13 before and after treatment, and is summarized in Table 14. Based on the results in Table 14 above, it can be seen that the reduction ratio of PASI before and after treatment with different biological agents is significantly related to the reduction ratio or the reduction difference of the ratio of interferon-γ/interleukin-13 in the patient, but the PASI reduction difference There was no significant correlation between the ratio of declines or the difference in declines in the patient's interferon-gamma/interleukin-13 ratio. Accordingly, the change in the expression of the ratio of interferon-γ/interleukin-13 before and after treatment (whether it is the decreasing ratio or the decreasing difference) can be used as a medication index for psoriasis patients.

Example 3 Screening drugs to treat people with psoriasis

Based on the results of Example 1 and Example 2, it was confirmed that the change in the expression level of interferon-γ and the ratio of interferon-γ/interleukin-13 before and after treatment can be used as an indicator of medication for psoriasis patients. The embodiment then evaluates the feasibility of the method of first screening drugs in vitro, that is, first using the cell screening platform of the present invention in vitro to screen out drugs suitable for the patient from which the cells are derived, and then administering the screened drug to the patient. drug.

Referring to Figure 1, the treatment results of a patient with psoriasis admitted to an outpatient clinic in this example (ie, patient C in Examples 1 and 2), the patient was initially treated with ^STELARA® after six months of treatment. , no significant improvement was found, the psoriasis lesions showed redness, the lesions were thick by visual inspection, and there was a lot of desquamation at close range (Panel A (back), Panel C (back of thigh), E Small image (back of calf (detail))). In order to seek the best therapeutic drug for the patient, the peripheral blood mononuclear cells of the patient are used to carry out the drug screening method of the present invention, and the drug that can produce the best curative effect on the patient is screened for treatment. It is found from the results of the screening method of the present invention that the peripheral blood mononuclear cells of the patient have a better response to ^HUMIRA® (the ratio of interferon-γ/interleukin-13 obtained after administration is ^HUMIRA® : 33.4, and STELARA ^® : 43.82). Accordingly, the follow-up treatment was switched to HUMIRA ^® , and after 6 months of treatment, it was found that the patient's condition showed significant improvement, the redness of the psoriasis lesions subsided, the thickness of the lesions did not thicken, and the desquamation decreased (Panel B (back), Panel D (back of thigh), Panel F (back of calf (partial)) in Figure 1), and the patient's PASI also improved from 13 before HUMIRA ^® treatment to 0.5 after ^HUMIRA® treatment.

Based on the above results, the drug screening method of the present invention firstly evaluates the changes in the expression of inflammatory cytokines secreted by cells in vitro (for example, the reduction ratio of the expression of interferon-γ, or the expression of interferon-γ/interleukin- 13), as the basis for the drug use of the individual from which the cells are derived, it can indeed achieve the purpose of significantly improving the efficacy of the drug, and has the benefit of applying for patent protection for inventions.

Although the above embodiments disclose specific embodiments of the present invention, they are not intended to limit the present invention. Those with ordinary knowledge in the technical field to which the present invention pertains, without departing from the principle and spirit of the present invention, should Various changes and modifications can be made to it, so the protection scope of the present invention should be defined by the appended claims.

none

The present disclosure and other features, aspects, and advantages will become more apparent upon review of the following detailed description, the scope of the claims, and the accompanying drawings, wherein:

Fig. 1 is a photographic diagram obtained according to an embodiment of the present disclosure, illustrating the change in the appearance of a patient's lesion before and after treatment after screening a therapeutic drug adalimumab (HUMIRA ^® ) by the method of the present disclosure, Among them, panels A, C, and E are before adalimumab treatment, while panels B, D, and F are after adalimumab treatment; panels A and B: back; panels C and D: posterior thigh ; E, F panels: the back of the lower leg (partial).

Claims (6)

A method for screening a therapeutic drug from a plurality of candidate drugs, wherein the therapeutic drug is used to treat psoriasis in an individual, comprising: (a) obtaining an isolated peripheral blood mononuclear cell from the individual cell, PBMC); (b) activate the peripheral blood mononuclear cells in step (a) with a stimulus, so that the cells produce interferon-gamma (interferon-gamma, IFN-gamma) and interleukin-13 (interleukin-13) , IL-13), wherein the stimulator is Streptococcus pyogenes ; (c) the activated peripheral blood mononuclear cells of step (b) are respectively administered the plurality of candidate drugs; (d) are detected separately The expression levels of the interferon-γ and the interleukin-13 in peripheral blood mononuclear cells in step (c); and (e) based on the interferon-γ and the interleukin detected in step (d) The expression level of -13, the therapeutic drug is screened from the plurality of candidate drugs, wherein the therapeutic drug has the highest inhibitory ability to the expression level of interferon-γ, or the expression level of interferon-γ is related to the interferon-γ expression level. The ratio of leukin-13 expression amount reached the minimum. The method of claim 1, wherein the therapeutic drug reduces the amount of interferon-gamma expression by at least 1% after administration of the candidate drug compared to before administration of the candidate drug. The method of claim 1, wherein the therapeutic drug reduces the expression of interferon-gamma and interleukin-13 by at least 1% after administration of the candidate drug compared to before administration of the candidate drug quantity ratio. The method of claim 1, wherein each of the plurality of drug candidates is selected from the group consisting of: Acitretin, alefacept, Anthralin, Apremilast ), Betamethasone, Calcipotriene, Calcipotriol, Calcitriol, Clobetasol, Coal tar, Cyclosporine ), Dithranol, Etanercept, Fluocinolone, Hydrocortisone, Infliximab, Methotrexate, Pimecromet Pimecrolimus, Tacrolimus, Tazarotene, Tretinoin, Bimekizumab, Brodalumab, Ixelizumab (Ixekizumab), Perakizumab, Remtolumab, Secukinumab, Vunakizumab, Brazikumab, Ancient Guselkumab, Mirikizumab, Risankizumab, Tildrakizumab, Ustekinumab, Adalimumab ), Certolizumab, Etanercept, Golimumab, Infliximab, and Lenercept. The method of claim 4, wherein the plurality of drug candidates are composed of: adalimumab, golimumab, guselkumab, ixelizumab, secukinumab, and ustekin composed of monoclonal antibodies. The method of claim 1, wherein the individual is a human.

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